Metabolic engineering of Escherichia coli to enhance hydrogen production from glycerol

Glycerol is an attractive carbon source for biofuel production since it is cheap and abundant due to the increasing demand for renewable and clean energy sources, which includes production of biodiesel. This research aims to enhance hydrogen production by Escherichia coli from glycerol by manipulati...

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Bibliographic Details
Published in:Applied microbiology and biotechnology 2014-05, Vol.98 (10), p.4757-4770
Main Authors: Tran, Kien Trung, Maeda, Toshinari, Wood, Thomas K
Format: Article
Language:English
Subjects:
biochemical pathways
biodiesel
Biodiesel fuels
Bioenergy and Biofuels
Biofuels
Biomedical and Life Sciences
Biotechnology
carbon
Dehydrogenases
E coli
Energy consumption
Enzymes
Escherichia coli
Escherichia coli - genetics
Escherichia coli - growth & development
Escherichia coli - metabolism
Ethanol
Fermentation
ferredoxin hydrogenase
Gene Knockout Techniques
genes
Genetic aspects
Genetic engineering
Glucose
Glycerin
Glycerol
Glycerol - metabolism
Hydrogen
Hydrogen - metabolism
hydrogen production
lactate dehydrogenase
Life Sciences
Metabolic Engineering
Metabolic Networks and Pathways
Metabolism
Microbial Genetics and Genomics
Microbiology
nitrate reductase
Oxidation
phosphoenolpyruvate carboxylase
Physiological aspects
Pollutants
pyruvic acid
Studies
Time Factors
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Summary:Glycerol is an attractive carbon source for biofuel production since it is cheap and abundant due to the increasing demand for renewable and clean energy sources, which includes production of biodiesel. This research aims to enhance hydrogen production by Escherichia coli from glycerol by manipulating its metabolic pathways via targeted deletions. Since our past strain, which had been engineered for producing hydrogen from glucose, was not suitable for producing hydrogen from glycerol, we rescreened 14 genes related to hydrogen production and glycerol metabolism. We found that 10 single knockouts are beneficial for enhanced hydrogen production from glycerol, namely, frdC (encoding for furmarate reductase), ldhA (lactate dehydrogenase), fdnG (formate dehydrogenase), ppc (phosphoenolpyruvate carboxylase), narG (nitrate reductase), focA (formate transporter), hyaB (the large subunit of hydrogenase 1), aceE (pyruvate dehydrogenase), mgsA (methylglyoxal synthase), and hycA (a regulator of the transcriptional regulator FhlA). On that basis, we created multiple knockout strains via successive P1 transductions. Simultaneous knockouts of frdC, ldhA, fdnG, ppc, narG, mgsA, and hycA created the best strain that produced 5-fold higher hydrogen and had a 5-fold higher hydrogen yield than the parent strain. The engineered strain also reached the theoretical maximum yield of 1 mol H₂/mol glycerol after 48 h. Under low partial pressure fermentation, the strain grew over 2-fold faster, indicating faster utilization of glycerol and production of hydrogen. By combining metabolic engineering and low partial pressure fermentation, hydrogen production from glycerol was enhanced significantly.
ISSN:0175-7598
1432-0614
DOI:10.1007/s00253-014-5600-3